除草剂莠去津降解菌SFAD3的分离鉴定及其土壤修复潜力

为获得能够修复除草剂莠去津污染土壤的高效降解菌,采用摇瓶富集法、平板分离法从莠去津过量使用的土壤中分离得到降解菌SFAD3,通过形态学、生理生化特征观察以及16S rDNA和ITS序列分析进行种类鉴定,测定获得菌株的最适降解条件,并通过土壤接种和盆栽试验验证菌株对莠去津污染土壤的修复作用。结果表明,菌株SFAD3最终被鉴定为门多萨假单胞菌Pseudomonas mendocina,该菌株培养30 d时对污染土壤中50 mg/kg莠去津的降解率可达72.6%;菌株SFAD3在MM液体培养基中最适降解条件为温度37℃、pH 7、莠去津初始浓度6.25 mg/L、接种量2%,对莠去津降的降解率为50.0%～72.2%;与仅有莠去津的处理相比,添加有SFAD3发酵液的处理20 d后芝麻的株高、根长、湿重和干重能够显著恢复,并且该菌对芝麻还具有一定的促生作用。表明降解菌SFAD3在修复莠去津污染土壤方面具有潜在的应用价值。     

猕猴桃溃疡病生防芽孢杆菌B2的鉴定及应用

猕猴桃溃疡病是一种毁灭性细菌病害,该病发展迅速,防治难度大,严重制约了猕猴桃产业的发展。为找到一种有效防治该病的方法,针对化学防治易产生抗药性、污染环境、杀伤天敌等特点,从猕猴桃根际土壤中分离共获得25株芽孢杆菌,采用点菌法筛选出4株对猕猴桃溃疡病菌(Pseudomonas syringae pv.actinidiae)具有拮抗效果的菌株(抑菌圈直径达到5 mm以上),其中B2拮抗作用最强。根据菌落(体)形态特征、生理生化鉴定以及16S r DNA序列分析,初步鉴定B2为蜡样芽孢杆菌(Bacillus cereus)。将B2的发酵原液稀释成1×109cfu/m L的发酵液,并在此基础上依次配制50、100、500、1000、2000倍的稀释液,用于盆栽试验。盆栽试验采用喷雾法,30天后的防治效果分别达到了90.45%、86.52%、80.90%、67.98%、47.19%、15.73%,60天后尽管病斑有少量增加,旧病斑有所增大,但与对照相比,病斑增长速度明显减缓。大田试验结果证明,该菌具有较好的生防潜力,喷雾法选择B2发酵液的50倍稀释液,涂抹法选择B2发酵液的100倍稀释液,可有效降低猕猴桃溃疡病的危害。     

Effect of biocontrol agents and biofertilizers on root rot,yield, harvest index and nutrient uptake of cassava (Manihot esculanta Crantz)

Cassava is an important subsidiary food and industrial raw material in the tropics. Root rot disease, caused by Phytophthora palmivora, poses a serious threat to cassava cultivation in Tamil Nadu, India. Field experiments (2008–09) were conducted to study the effect of biocontrol agents (Trichoderma spp. and Pseudomonas fluorescens) and biofertilizers (Azospirillum, vesicular–arbuscular mycorrhizal fungi and phosphorus-solubilizing bacteria) on root rot, yield, harvest index and nutrient uptake of cassava at two NPK rates. The design of the experiment was a split plot with two NPK rates, recommended and 50% recommended rate, as the main plot treatments and five biocontrol agents and biofertilizers as subplot treatments. The results clearly indicated that use of a bioinoculants consortium significantly reduced root rot infection/disease incidence over uninoculated controls. Azospirillum significantly improved the yield of cassava at 50% of the recommended rate of NPK. NPK rates had no significant impact on harvest index of cassava and Trichoderma and vesicular–arbuscular mycorrhizal fungi resulted in a higher harvest index even at 50% of the recommended NPK rate. Nitrogen, phosphorus and potassium uptake was significantly improved when treated with biofertilizers and/or a consortium.     

A New Cell Wall Located N-rich Protein is Strongly Induced During the Hypersensitive Response in Glycine Max L.

Soybean (Glycine max (L.) Merill, cv. Williams 82) plants and cell cultures respond to avirulent pathogens with a hypersensitive reaction. After inoculation of soybean with Pseudomonas syringae pv. glycinea, carrying the avirulence gene avrA, or zoospores from the fungus Phytophthora sojae Race 1, a resistance-gene-dependent cell death programme is activated. A new gene was identified by differential display of mRNAs that is specifically activated during the early phase of incompatible pathogen-soybean interactions but does not respond to compatible pathogens. The gene is strongly induced within 2h after addition of P. sojae zoospores. A similar kinetic pattern was observed for P. syringae (avrA) inoculated soybean cell cultures. The gene encodes a deduced protein of 368 amino acids with a very high content of asparagine and was therefore termed N-rich protein (NRP). The protein is composed of two distinct domains, of which only the C-terminal domain has striking homology to proteins of unknown function from other plants. An antibody raised against the recombinant NRP recognizes a protein of 42kDa. The protein is located in the cell wall as indicated by cell fractionation studies. Comparison of the genomic DNA-sequence with the cDNA, identified two introns within the open reading frame. The NRP-gene is not directly induced by salicylic acid or hydrogen peroxide, indicating a distinct and specific signal transduction pathway which is only activated during programmed cell death. The NRP-gene appears to be a new marker in soybean activated early in plant disease resistance.     

Control of Phytophthora cryptogea in the hydroponic forcing of witloof chicory with the rhamnolipid-based biosurfactant formulation PRO1

Rhamnolipids, extracellular metabolites of Pseudomonas aeruginosa with surfactant properties, proved to be very effective in controlling the spread of brown root rot disease caused by Phytophthora cryptogea in the hydroponic forcing system of witloof chicory ( Cichorium intybus var. foliosum ). The biosurfactant was applied as the product PRO1, a formulation of 25% rhamnolipids in oil. Both an in vitro screening and in vivo experiments in a mini-hydroponic system demonstrated the ability of PRO1 to control brown root rot. A 25  µ g mL⁻¹ rhamnolipids nutrient solution was enough to obtain good control of an artificial infection with a zoospore suspension of P. cryptogea . The biosurfactant PRO1 performed well in a semicommercial system under growers' conditions. A treatment of 25  µ g mL⁻¹ rhamnolipids (100  µ g mL⁻¹ PRO1) reduced the disease incidence significantly in two independent experiments. However, PRO1 was not effective when a mycelial suspension was used as inoculum. Rhamnolipids have good potential to limit the spread of P. cryptogea in the hydroponic forcing system of witloof chicory, and can be used as a preventive measure against brown root rot.     

纳米TiO2悬浮液对烟草青枯菌杀菌活性的研究

纳米T iO2是一种半导体光催化剂,在近紫外光的激发下,可以产生强的杀菌活性。以烟草青枯菌P seudomonas solanacearum为试验对象,采用平板稀释法对自制的纳米T iO2悬浮液进行杀菌活性测定。结果表明:在阳光或汞灯下照射4h后,T iO2对烟草青枯菌的杀菌率可达90%以上。说明用纳米T iO2防治烟草青枯病具有一定的可行性。     

Characterization of Elicitor-inducible Tobacco Genes Isolated by Differential Hybridization

Inducible responses in plants against pathogen attack play a major role in resistance to disease. The defense responses are mostly associated with the expression of various kinds of inducible genes. We employed differential hybridization to isolate elicitor-inducible genes (EIGs) of tobacco (Nicotiana tabacum cv. Samsun NN) using the tobacco-fungal elicitor system. A cDNA library was constructed from tobacco leaves treated for 12 hr with hyphal wall components (HWC) prepared from Phytophthora infestans, and six EIGs were identified. Expression of all EIGs was induced after inoculation with the soybean pathogen Pseudomonas syringae pv. glycinea (nonpathogenic on tobacco) or treatment with salicylic acid, and a variety of expression patterns of EIG mRNAs was observed. Sequence analysis of EIG cDNAs revealed similarities to genes for SAR8.2 (EIG-B39 and EIG-D14), glycine-rich protein (EIG-G7), extensin (EIG-I30), acyltransferase (EIG-I24) and unknown protein (EIG-J7). Possible roles of EIG products in disease resistance are discussed. Received 30 August 2000/ Accepted in revised form 30 November 2000     

抑菌圈-定殖力双重测定法筛选青枯病生防细菌

 本研究首先用平皿抑菌圈法筛选出55个拮抗青枯菌的细菌菌株,将番茄幼苗在各菌菌悬液中浸根12h后栽种于温室未灭菌的土壤中,结果发现,有22个菌株在幼苗根部的定殖能力较强(终定殖密度大于104 cfu/g根),其中革兰氏阴性土壤细菌占同类菌的86.3%,革兰氏阳性土壤细菌占同类菌的13.0%,10个无致病力产细菌素的青枯菌菌株在根部的终定殖密度均低于104 cfu/g根,其定殖能力弱于致病青枯菌。有17个拮抗菌菌株在番茄幼苗根部的定殖密度超过所有致病菌。温室生防结果证明,抑菌圈-定殖力双重测定法对于筛选生防菌株是一种行之有效的方法。     

荧光93菌体和代谢物对全蚀病菌和小麦生长影响的初步研究

对荧光假单胞菌工程菌剂“荧光93”菌体和代谢物的作用特性进行了比较。结果表明:该菌剂能保护小麦根系免遭全蚀病危害,对植株生长有促进作用。菌体的抑菌能力强于代谢物,浸种处理防效分别为44.8％和27.8％;代谢物具有更强的刺激植株生长作用,在无病条件下,代谢物灌根处理鲜重增加34.5％,菌体处理鲜重增加7.5％。培养基成份对代谢物的作用有一定影响:以马铃薯葡萄糖培养产生的代谢物,抑菌效果好;营养肉汁酵母粉培养产生的代谢物,促生作用好。据此推断,荧光93菌剂代谢产物中含有抑制全蚀病菌和刺激植株生长的物质。     

Need for flagella for complete virulence of Pseudomonas syringae pv. tabaci: genetic analysis with flagella-defective mutants ?fliC and ?fliD in host tobacco plants

The flagellins purified from Pseudomonas syringae pv. tabaci induce a hypersensitive reaction in nonhost tomato cells. To investigate the role of flagella and flagellin in the compatible interaction, we generated two types of flagella-defective mutant. The fliC mutant lost the fliC gene that encodes flagellin protein, whereas the fliD mutant lost the fliD gene that encodes HAP2-capping protein. The two mutants had markedly reduced ability to cause disease symptoms in tobacco leaves. Furthermore, propagation of the mutants in tobacco leaves was less than that in wild-type pv. tabaci. Compared to the inoculation with wild-type pv. tabaci, inoculation with the two mutants did not markedly induce the expression of typical defense response-related genes such as PAL and hsr203J. Complementation of each fliC and fliD gene to the corresponding deficient mutant restored motility and virulence. These results indicate that flagella of P. syringae pv. tabaci are indispensable organelles for complete virulence on host tobacco plants.